The main advantage of HPLC (high-pressure liquid chromatography; High-performance liquid chromatography) purification is the monitoring of the purity level. It is recommended for applications such as diagnostic primers and antisense probes.
Reverse-Phase HPLC (RP-HPLC) operates on the same principle as RP-cartridge-Gold™ purification. Full-length oligonucleotide (containing DMT group at the 5’ OH of the synthetic oligonucleotide last base) established hydrophobic interaction with the alkyl chains bonded covalently to the support surface and was then retained on the matrix while truncated forms passed through. Due to the inherent hydrophobicity and non-specifically binding on the matrix of long oligonucleotides, RP-HPLC can be used to purify oligonucleotides up to 60 nucleotides in length but remains a method of choice to purify larger amounts of oligonucleotides (> 1000 nmol) with a purity level up to 85%.